We propose to produce human monoclonal antibodies to cytomegalovirus (CMV) as a potential unlimited and safe source of antigen-specific antibodies and to analyze their viral antigen specificity and functional effects on the immune response to virus in normal and immunocompromised individuals. While impaired cellular immunity is thought to be the major contributing factor in immunocompromised patients, administration of human plasma with high anti-CMB titers modifies with severity or eliminate CMV infections. Among bone marrow and organ transplant recipients, reactive or acquired herpesvirus infections frequently occur after transplantation, with blood or blood component transfusions as a significant vector in virus acquisition. (CMV) is the herpesvirus most commonly associated with significant morbidity and mortality. Normal individuals, when challenged with a viral infection, generate cytotoxic lymphoid and non-lymphoid killer cells, as well as viral specific antibody which can neutralize virus directly, as well as provide a """"""""bridge"""""""" between lymphoid killer cells and infected target cells in antibody-dependent cellular cytotoxicity. A human-mouse derived myeloma line, shown to fuse efficiently to human B cells and produce stable human immunoglobulin secreting hybridomas, will be fused to B cells of anti-CMV seropositive patients with demonstrable in vitro secretion of anti-CMV antibodies to produce a panel of human monoclonal antibody against CMV. Using this system, several human monoclonal antibodies against CMV have already been produced. The effects of these antibodies and others to be produced on antigen specific immune function will be explored by adding antibody to in vitro assays. Finally, the target molecules of these antibodies will be radiolabeled in virus infectioned cells and analyzed structurally by polyacrylamide gel electrophoresis. These studies should provide new insights into the basic mechanisms underlying the immune response to viral infections. These antibodies should also be useful in the diagnosis and treatment of CMV infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022557-02
Application #
3133801
Study Section
Experimental Virology Study Section (EVR)
Project Start
1985-09-30
Project End
1987-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Bradshaw, P A; Duran-Guarino, M R; Perkins, S et al. (1994) Localization of antigenic sites on human cytomegalovirus virion structural proteins encoded by UL48 and UL56. Virology 205:321-8
Perkins, S; Zimmermann, U; Foung, S K (1991) Parameters to enhance human hybridoma formation with hypoosmolar electrofusion. Hum Antibodies Hybridomas 2:155-9
Foung, S; Perkins, S; Kafadar, K et al. (1990) Development of microfusion techniques to generate human hybridomas. J Immunol Methods 134:35-42
Zimmermann, U; Gessner, P; Schnettler, R et al. (1990) Efficient hybridization of mouse-human cell lines by means of hypo-osmolar electrofusion. J Immunol Methods 134:43-50
Foung, S K; Perkins, S; Bradshaw, P et al. (1989) Human monoclonal antibodies to human cytomegalovirus. J Infect Dis 159:436-43
Foung, S K; Perkins, S (1989) Electric field-induced cell fusion and human monoclonal antibodies. J Immunol Methods 116:117-22